PCR core kit with hot-start function and proofreading function for high sensitivity and accurate amplification
• High accuracy, specificity and low error rate
– Added Hot-start function of i-StarTaq ™ DNA polymerase
– Proof-reading activity enables amplification of long DNA template
– High amplification efficiency and sensitivity to amplification of long and short template DNA
• Applicable to various DNA from cloned DNA, human genomic DNA to templates that are difficult
The i-StarMAX ™ II DNA Polymerase has a hot-start function, which significantly reduces nonspecific reactions. It is very useful for PCR of long templates of 20 Kb or less. Since Taq DNA Polymerase has good activity even at 37 ℃, occasionally, the primer is attached to the DNA molecule and extension occurs before the first denaturation process is completely progressed. If annealing occurs at such a low temperature, there is a high probability that the primer will mismatch, resulting in a band with poor accuracy. Forward or reverse primer is either specific or the two primers are prone to generate a primer dimer. This phenomenon occurs when the Taq DNA Polymerase is activated only at high temperature by hot-start PCR and the PCR specificity and sensitivity It is to use to raise. In Hot start PCR, the activity of primer (polymerase, MgCl2, dNTP) is blocked so that the primer can be attached to the desired DNA site exactly, and when the temperature of the reaction becomes high, it becomes active, thereby preventing nonspecific priming or oligomerization I will. This avoids mismatching as described above, so you get a relatively clear band. As described above, i-StarMAX ™ II DNA Polymerase is a mixture of hot-start i-StarTaq ™ DNA Polymerase and Proofreading DNA Polymerase. It reduces nonspecific reaction and error rate and PCR a template longer than 20 kb Very useful. It is also useful for demanding template / primer systems. High yield PCR products can be obtained by setting the extension time of PCR to 30 ~ 60 seconds per kb. i-StarMAX ™ DNA Polymerase is suitable for high-performance amplification and rapid PCR of 15 Kb or more of PCR targets that are difficult to achieve with proofreading DNA polymerase alone. Most of the products amplified with this enzyme have one more base A at the 3 ‘end. Therefore, the PCR product can be cloned into a T-vector as well as cloned into a blunt end by cloning into DNA polymerase such as Klenow DNA polymerase or T4 DNA polymerase, and cloned into a blunt-end vector through phosphorylation.
Amplification efficiency observation from Hot-Start PCR
i-StarMAX ™ II DNA polymerase has been confirmed for various template and size sensitive activity. We have confirmed that the sensitivity and amplification efficiency are higher than those of other hot-start Taq DNA Polymerase.
Lane M, 100 bp Ladder DNA Marker; lane 1, 100 ng genomic DNA; lane 2, 5-1 diluted genomic DNA; lane 3, 5-2 diluted genomic DNA; lane 3, 5-3 diluted genomic DNA; lane 4, 5-4 diluted genomic DNA; lane 5, 5-5 diluted genomic DNA; lane 6, 5-6 diluted genomic DNA; lane 7, 5-7 diluted genomic DNA
Sensitivity comparison data
In order to measure the sensitivity of DNA amplification, we compared i-StarMAX™ II DNA Polymerase with products of the same function. As can be seen from the results, i-StarMAX ™ II DNA Polymerase is effectively amplified from a small amount of DNA compared to other products.
Lane M1, 100 bp Ladder DNA Marker; lane 1, 25 ng gDNA; lane 2, 5 ng gDNA; lane 3, 1 ng gDNA; lane 4, 200 pg gDNA; lane 5, 40 pg gDNA; lane 6, 8 pg gDNA